15 Responses to “* Excerpt from a Representative Rabbit Protocol Involving Intradermal Challenge By Bruce Ivins (such as he was doing in early October 2001)”

DXersaid

You may recall that Flask 1029 was purified with Renocal. An alternative to purifying with renocal is through heat-shocking. Heat shocking is done in Building 1412 in connection with aerosol experiments — where animals were challenged with a wet aerosol. Here is an experiment on which Dr. Ivins was principal investigator involving a study of heat shocked vs. non-heat shocked spores.

DXersaid

It’s time to upload a copy of an unredacted copy of B01-11, the animal protocol relating to the formaldehyde/ rabbit experiment that Dr. Ivins was working in late September 2001 and early October 2001 when the AUSAs speculated Dr. Ivins was powderizing anthrax that was then used to murder people.

DXersaid

In this 1998 protocol involving intradermal challenge of rabbits, which can be found in a lab notebook at the USAMRIID FOIA room, the protocol only involves the rabbits being checked 2Xs a day, not 3X.

If Ivins had contrived this as his alibi then he would have remembered it and used it before he lost his security clearance. The fact that Ivins forgot this and its rich history of documentary evidence shows that in fact he could not remember what he did those nights.

That shows he didn’t prepare anthrax on those nights. Because if he had, he would have remembered an alibi he so carefully constructed a paper trail for. Instead, he never raised it before losing his clearance, being fired, having people told not to talk to him, and taking his life.

“Even if there is appropriate airflow from the office to the suite hallways, these configurations pose significant risk to laboratory personnel, since it is likely that contamination can easily occur and, in fact, was documented in B303.”

Returning to the bacteremia, where was the blood infected with anthrax cultured? The BL-3? Rm. 313? Dr. James Baker once emphasized to me that such work with infected animals (challenged subcutaneously) needed to be done in a BL-3 which is why it was done by Bruce Ivins at USAMRIID and not at University of Michigan.

DXersaid

3/25/2004 302 Interview statement
–
Dr. Ivins did not have his emails from 2001 so as to be able recreate events.

“IVINS consulted with _____________ at USAMRIID Computer Services and learned that electronic mail (email) can be retrieved for a two year period, however it is expensive to do so. If more than two years have passed, it is not possible to retrieve email.”

DXersaid

If he had been walked through his emails, he could have discussed his correspondence in the summer of 2001 in which someone was having problems with foaming and Dr. Ivins explained that they did not use an antifoaming agent in their experiments (because it might mess up the vaccine work by introducing a variable). (Antifoaming agents, e.g., aerosil, commonly are silica-based btw). He also separately offers to “drive down” the vaccine needed for one challenge.

From: Ivins, Bruce E Dr USAMRIID
To:
Bcc:
Subject: RE: Spores and Foaming
Date: Tuesday, June 05, 2001 7:29:36 AM
When we mix the spores at that concentration, we don’t vortex. I should have said that. I think the
reason that it may foam is that the spore suspension is so pure. In the Vollum 1B spore suspension from
1965 which is about 4 – 5 X 10^10 per ml, and MUCH dirtier, there is no foaming upon vortexing. The
spores you have were twice purified with Hypaque gradient centrifugation. The spores are very
hydrophobic, I believe. I suppose you could try to add a little Tween 80 to the spores to see if that
helps. I’ve heard that in the “OLD DAYS” back in the 50s and 60s here at Detrick, they would soemtimes
add a little Tween 80 to the spores to be aerosolized. We haven’t added any because we didn’t want to introduce another variable into the challenge. If you add something else to the spores being aerosolized,

you may have to be able to demonstrate that the “anti-foam” has no effect on spore LD50, the infection
process, or the specific immune response to the infection. As I said, when we mix the spores at that
concentration, we just rock back and forth or gently swirl. If you want to add something to the spores
before challenge, I think you should first run it by the IPT for their comments.
– Bruce

—–Original Message—–
From:
Sent: Monday, June 04, 2001 5:06 PM
To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’
Subject: Spores and Foaming
Bruce,
thought it would be easier if I contacted you directly. With regard to
the foaming issue. When I go back to the original suspension you sent in
the 50 ml conical tube and vortex it the same foaming phenomenon occurs. So
I do not believe it is a glassware problem or washing problem. If you
will/could go back to one of your 10E10 stocks of the same spore prep. and
also make a 10E9 dilution and vortex it to see if you get the same thing.
As described before, it’s like whipped cream on top of the water and
will not go back into suspension unless it sits for a day or more. When I
enumerate the suspension under the whipped cream it is 0.5-1 log lower than
what is expected (i.e. what should be 10E9 is 5 x 10E8 to 1 x 10E8). In the
mean time do you have any ideas on a defoaming agent?

From: Ivins, Bruce E Dr USAMRIID
To:
Bcc:
Subject: RE: Spores and Foaming
Date: Tuesday, June 05, 2001 9:04:29 AM
, these spores are exactly the same spores used for the other rabbits for BioPort. We do get some
foaming, but still get a high dose with 3 X 10^9 per ml. Would it help to use more suspension in your
container? We’ve used these spores for quite awhile with success. As I said, maybe it’s because they
are so clean that they clump. If there were some cell material stuck to the outside, perhaps they would
perform a little more like the old Vollum 1B spore suspension or like some of the less pure Ames spore
suspensions we have used in the past.
– Bruce

—–Original Message—–
From:
Sent: Tuesday, June 05, 2001 8:24 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Cc:
Subject: RE: Spores and Foaming
Bruce,
One other question. Is were the spores that you sent to us prepared the
same way as the ones RIID used on the BioPort rabbit studies or the same
spore prep.? Or did you use different AMES spores? Looks like even though
I’m a log low on the AGIs than expected I can still hit the targeted LD50
range and will use These spores and mix by inversion. Thanks for answering
my questions

—–Original Message—–
From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Tuesday, June 05, 2001 7:30 AM
To: ‘
Subject: RE: Spores and Foaming
, When we mix the spores at that concentration, we don’t vortex. I should
have said that. I think the reason that it may foam is that the spore
suspension is so pure. In the Vollum 1B spore suspension from 1965 which is
about 4 – 5 X 10^10 per ml, and MUCH dirtier, there is no foaming upon
vortexing. The spores you have were twice purified with Hypaque gradient
centrifugation. The spores are very hydrophobic, I believe. I suppose you
could try to add a little Tween 80 to the spores to see if that helps. I’ve
heard that in the “OLD DAYS” back in the 50s and 60s here at Detrick, they
would soemtimes add a little Tween 80 to the spores to be aerosolized. We
haven’t added any because we didn’t want to introduce another variable into
the challenge. If you add something else to the spores being aerosolized,
you may have to be able to demonstrate that the “anti-foam” has no effect on
spore LD50, the infection process, or the specific immune response to the
infection. As I said, when we mix the spores at that concentration, we just
rock back and forth or gently swirl. If you want to add something to the
spores before challenge, I think you should first run it by the IPT for
their comments.
– Bruce

From: Ivins, Bruce E Dr USAMRIID
To:
Bcc:
Subject: RE: Spores and Foaming
Date: Tuesday, June 05, 2001 9:20:04 AM
,
We usually spray at a concentration of 3 X 10^9 per ml. That gives us an aerosol inhaled dose of about
100- 200 LD50s in a 10-minute spray. You can try a test run with some Tween 80 and see if that helps.
I seem to recall they used some concentration between 0.01% and 1%, but I don’t remember exactly,
since it was given to me by word-of-mouth. I still recommend getting the IPT’s opinion. If there’s no
other way to aerosolize than using anti-foam, you may have to do so, but I would hesitate to do it
unless absolutely necessary.
– Bruce

—–Original Message—–
From:
Sent: Tuesday, June 05, 2001 8:02 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Spores and Foaming
True but when I nebulize a 10E9 conc. the foaming happens. Do you not
nebulize that high of a conc.? Also even at lower dilutions my AGI
enumerations are approx. 1 log lower than what I expect. Thus I appears
that even al low conc. they foam out of suspension and I’ll have to add some
type of defoaming agent.

From:
Sent: Tuesday, June 05, 2001 9:32 AM
To: ‘
Cc: ‘bruce.ivins@det.amedd.army.mil’
Subject: RE: Spores and Foaming
: I believe we are resolving our questions regarding the foaming and
we won’t be vortexing anymore. Bruce has helped us out immensely (see
below). Could you please provide information regarding the anti-foam
formulation that your staff uses – if any – for these high concentration
anthrax aerosols.
Thanks

—–Original Message—–
From:
Sent: Tuesday, June 05, 2001 9:22 AM
To:
Subject: FW: Spores and Foaming
—–Original Message—–
From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Tuesday, June 05, 2001 9:20 AM
To: ‘
Subject: RE: Spores and Foaming
We usually spray at a concentration of 3 X 10^9 per ml. That gives us an
aerosol inhaled dose of about 100- 200 LD50s in a 10-minute spray. You can
try a test run with some Tween 80 and see if that helps. I seem to recall
they used some concentration between 0.01% and 1%, but I don’t remember
exactly, since it was given to me by word-of-mouth. I still recommend
getting the IPT’s opinion. If there’s no other way to aerosolize than using
anti-foam, you may have to do so, but I would hesitate to do it unless
absolutely necessary.
– Bruce

—–Original Message—–
From:
Sent: Tuesday, June 05, 2001 8:02 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Spores and Foaming
True but when I nebulize a 10E9 conc. the foaming happens. Do you not
nebulize that high of a conc.? Also even at lower dilutions my AGI
enumerations are approx. 1 log lower than what I expect. Thus I appears
that even al low conc. they foam out of suspension and I’ll have to add some
type of defoaming agent.

From:
Sent: Tuesday, June 05, 2001 9:57 AM
To: Ivins, Bruce E Dr USAMRIID
Cc:
Subject: RE: Spores and Foaming
We need to look at your spray factor and adjust accordingly – we do NOT want to change anything
from what we do here – I know Bruce is being helpful – BUT——
Can I see the numbers and starting conc. etc.????
Thanks,

–

From:
Sent: Tuesday, June 05, 2001 8:02 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Spores and Foaming
True but when I nebulize a 10E9 conc. the foaming happens. Do you not
nebulize that high of a conc.? Also even at lower dilutions my AGI
enumerations are approx. 1 log lower than what I expect. Thus I appears
that even al low conc. they foam out of suspension and I’ll have to add some
type of defoaming agent.

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: Spores
Date: Tuesday, June 05, 2001 1:49:25 PM
Attachments:
Hi,
I reworded the Statement of Work for the anthrax spores according to what you said. I’ve
enclosed the file here. Please let me know if this looks OK. When it meets your OK, I’ll send it over to
Thanks, and see you in Annapolis!!
– Bruce

—–Original Message—–
From:
Sent: Wednesday, June 06, 2001 12:08 PM
To:
Cc: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Spray factor data
Enclosed is a representative sample of what I typically see. Are these
spray factors more in line with what you see or are they still not as
efficient? Some time during the conference can you, myself and
get together to discuss this? Also, thought it might be worth
while if after the conference on Wed. or the IPT on Friday if it might be
possible for me to come to USAMRIID and see if your spores look similar
(they should) and react the same why after making a 10E9 suspension and
vortexing and a 10E9 suspension then nebulizing to see if the foaming
occurs. Assuming you have the time and it is allowable. As you saw from
the spay factor calc. you determined, I can not achieve the targeted aerosol
conc. to reach 100-200 LD50s for the BioPort study.

—–Original Message—–
From:
Sent: Wednesday, June 06, 2001 10:23 AM
To: ‘
Subject: RE: Spray factor data
Attached to spread sheet is my recalculated spray factors – the way we
calculate etc.
By my calculations – an aerosol concentration of around 5 X 10(6) cfu/l is
needed for the animals to get around 150 LD50s
Your spray factors are in the low range compared to ours – we usually get
better efficiency
Are you going to repeat this???

—–Original Message—–
From:
Sent: Tuesday, June 05, 2001 4:24 PM
To:
Cc:
Subject: Spray factor data
Here is the spray factor information that you have been waiting for.
Limited reps. because we had the foaming problem. It looks like I would
have to start with a neb. conc. of approx. 2.9 X10E9 to hit 100-200 LD50s.
Do you usually a larger drop in AGI conc. from the initial neb. 10E9
concentration as compared to the lower dilutions?

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: RE: – Stability Indicating Assay sub-team
Date: Thursday, June 07, 2001 8:06:45 AM
Hi,
telephone number and email address are:
I think that you will find him a very competent and knowledgeable individual, with a
great deal of personal experience with respect to fermentation, production of antigen, adsorption onto
Alhydrogel, analysis of antigen product, and desorption from Alhydrogel.
Sincerely,
Bruce Ivins

—–Original Message—–
From:
Sent: Thursday, June 07, 2001 7:52 AM
To: ‘Bruce.Ivins@DET.AMEDD.ARMY.MIL’
Subject: – Stability Indicating Assay sub-team
Dear Bruce,
I am wondering if you could provide me with telephone number and his
E:mail address so that I can contact him. I do not have USAMRIID directory
with me.
Regards,

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: RE: Suggestions for study
Date: Wednesday, June 13, 2001 5:21:39 PM
Sure, When would you like the material? How many doses of each? Should I drive it down to
you, or is there someone here that can get it to you?
Regards,
– Bruce

—–Original Message—–
From:
Sent: Monday, June 11, 2001 11:31 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
The monkeys are arriving for the anthrax study. Any chance I can get the
vaccines (both the new and the old) from you to start the immunizations?

—–Original Message—–
From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Thursday, May 31, 2001 3:18 PM
To:
Subject: RE: Suggestions for study
I forgot to add that I wasn’t sure of the final approved groups for
vaccination, and if some of the groups received reduced levels of PA.
– Bruce

—–Original Message—–
From:
Sent: Thursday, May 31, 2001 2:32 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
The ACUC has approved our anthrax vaccine study in rhesus macaques. We
should be getting the animals in within a month. We have approval to test
the effect of our CpG ODN on both the old and new vaccines, if you can
provide them for immunization.
Hooray.

—–Original Message—–
From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Friday, May 11, 2001 7:58 AM
To:
Subject: RE: Suggestions for study
Hi,
I’ll be happy to provide you with the information. The current,
FDA-approved human anthrax vaccine consists of supernatant material,
primarily anthrax protective antigen (in undetermined and varying amounts),
from fermentor cultures of a toxinogenic, non-encapsulated strain of B.
anthracis, V770-NP1-R. Each 0.5-ml dose of the vaccine delivers about 0.725
mg of metallic aluminum (from the aluminum hydroxide adjuvant), protective
antigen in the range of about 0.5 to 20 micrograms, and other material such
as lethal factor, some edema factor (in certain lots but not others) and
other cellular material. The proposed new vaccine will contain less aluminum
(0.5 mg/dose), no lethal factor, no edema factor, and no other B. anthracis
material other than a specified and constant, defined amount of protective
antigen. We are tentatively targeting 50 micrograms as that amount. Use of
the proposed new vaccine in rabbits and rhesus macaques has demonstrated
efficacy against challenge, but has not demonstrated any observable
morbidity, mortality, or local or systemic reactogenicity. (Formal toxicity
studies are scheduled, but have not been conducted yet.) Thus, there is good
evidence of protection, but no evidence of adverse reactions associated with
the product. I should point out that the original vaccine contains
formaldehyde, which may contribute to some of the reactogenicity seen in
humans with the current vaccine. The vaccine you will be testing will not
contain formaldehyde. We have not yet published our findings with respect to
toxicity/reactogenicity of the new vaccine. Unfortunately, the studies
describing efficacy of the new vaccine are in abstract form, but have yet to
be put into a formal publication. I would suggest the following references
are pertinent to the concerns of your ACUC:
1. Comparative efficacy of experimental anthrax vaccine candidates
against inhalation anthrax in rhesus macaques. B. E. Ivins, M. L. M. Pitt,
P. F. Fellows, et al. 1998. Vaccine 16:1141-1148.
2. Comparative efficacy of a recombinant protective antigen vaccine
against inhalation anthrax in guinea pigs, rabbits, and rhesus monkeys. M.
L. M. Pitt, B. Ivins, J. E. Estep, et al. 1996 Abstracts of the Annual
Meeting of the American Society for Microbiology. E-70, p. 278.
3. Comparison of the efficacy of purified protective antigen and
MDPH to protect non-human primates from inhalation anthrax. M. L. M. Pitt,
B. E. Ivins, J. Farchaus and A. M. Friedlander. 1996. Salisbury Medical
Bulletin Special Supplement, # 87, p. 130.
I hope this has been helpful.
– Best regards,
– Bruce
—–Original Message—–

From:
Sent: Thursday, May 10, 2001 3:54 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
Hope all is well.
Our protocol was reviewed this morning by the ACUC. They’ve tentatively
approve it, with the caveat that I need to provide more background on the
safety of the vaccine immunogen. Specifically, they want to know about the
PA-based vaccine as well as the original vaccine (which I hope to include as
a positive control). What can you tell me about the safety of these agents?
Can you provide some background on how they are manufactured, how pure they
are, and what other studies have been done that support going into monkeys?
All the best,
—–Original Message—–

From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Tuesday, April 17, 2001 7:26 PM
To:
Subject: RE: Suggestions for study
Yes, I can get you both the current vaccine and the new candidate
vaccine. Please email the protocol when it is in final form. Also, please
let me know what you’ll need, when you’ll need it, and how much you’ll need.
Also, We were going to do ELISAs and TNA assays. The mouse passive
protection may be a bit dicey as far as getting conclusive results, since
mice aren’t a very good model for anthrax and specific protection against
anthrax. Good luck on the protocol.
– Bruce

—–Original Message—–
From:
Sent: Tuesday, April 17, 2001 1:29 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
I submitted the ACUC protocol this morning. Wish us luck. Do you have
access to both the PA vaccine and the old approved vaccine? I’m wondering
whether it would make more sense to study 5 monkeys/group, and immunize
different animals with one or the other type of vaccine +/- CpG ODN. Since
you’re transferring the serum to mice, and can study a large number of mice
per donor monkey, this may allow us to precisely characterize which vaccine
is better, and whether CpG’s work.
What do you think?
—–Original Message—–

From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Wednesday, April 11, 2001 1:18 PM
To:
Subject: Suggestions for study
Hi,
I’m including a Word document with suggestions for the CpG/monkey
study. Please feel free to modify it as much as you wish. If you have any
questions, please ask them, either by email or phone.
– Bruce

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: RE: Suggestions for study
Date: Wednesday, June 13, 2001 5:44:16 PM
,
We usually make up the vaccine for use no more than about 2-3 days ahead of schedule,
although we’ve not done the stability studies to see whether we can go longer or not. (My guess is that
we could. I just don’t.) Rather than ship it, I would rather carry it down to you on gel ice in person.
That way, nothing could happen en route with the third-party shipper/handler. I could get it down to
you the first part of next week if needed, or later if desired. I’ll get you extra amounts of each vaccine.
It’s no problem to make the vaccine, and it will only take a couple of hours to drive down, give it to
you, and come back. I’m quite excited about the experiments.
– Bruce

From:
Sent: Wednesday, June 13, 2001 5:37 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
I believe the protocol calls for a prime/boost, just as planned for use in
humans. We will have 10 animals/arm, and thus need 20 doses each of the new
vaccine and the old vaccine. As I recall, they are already in alum, ready
for administration. We’ll just add our ODN and go. Naturally, we need a
bit extra since there’s some wastage.
In terms of timing, that depends on how stable the vaccine is. I assume its
made up in advance and stored in the fridge? If so, we could accept it
immediately, and start the injections as soon as we can get on the animal
handler’s schedule. If it’s perishable, we’ll have to plan 5the timing and
then let you know. In terms of getting it here, can you ship it on ice?
Seems a shame to make you drive all the way down. If you or a colleague are
coming this way, we’d be happy to wait or a hand delivery.
Let me know, or perhaps we should chat by phone?
—–Original Message—–

From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Wednesday, June 13, 2001 5:22 PM
To: ‘
Subject: RE: Suggestions for study
Sure, When would you like the material? How many doses of each?
Should I drive it down to you, or is there someone here that can get it to
you?
Regards,
– Bruce

—–Original Message—–
From:
Sent: Monday, June 11, 2001 11:31 AM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
The monkeys are arriving for the anthrax study. Any chance I can get the
vaccines (both the new and the old) from you to start the immunizations?
—–Original Message—–

From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Thursday, May 31, 2001 3:18 PM
To:
Subject: RE: Suggestions for study
I forgot to add that I wasn’t sure of the final approved groups for
vaccination, and if some of the groups received reduced levels of PA.
– Bruce
—–Original Message—–

From:
Sent: Thursday, May 31, 2001 2:32 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Suggestions for study
Dear Bruce,
The ACUC has approved our anthrax vaccine study in rhesus macaques. We
should be getting the animals in within a month. We have approval to test
the effect of our CpG ODN on both the old and new vaccines, if you can
provide them for immunization.
Hooray.

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: Thanks again!
Date: Thursday, June 14, 2001 6:17:29 AM
I just wanted to tell you once again how much we appreciate all your efforts on the 4th
International Conference on Anthrax. We enjoyed working with you both immensely. The comments that
we heard from many other attendees point to the meeting having been a great success, in large part
due to you. You are both very competent and very personable, and you are a credit to the ASM. Take
care, and many thanks again!
Sincerely,
Bruce Ivins
Research Microbiologist
USAMRIID Bacteriology Division

From: Ivins, Bruce E Dr USAMRIID
To: Ivins, Bruce E Dr USAMRIID
Cc:
Subject: RE: Spores and Foaming
Date: Friday, June 15, 2001 12:16:15 PM
,
I am sending you on Monday, 3 15-ml polypropylene tubes of Ames spores, each of which
contains about 10-11 ml of spores at 3.9 X 10^10 per ml. Here are suggestions as to how to handle
them to minimize foaming. This is how we handle them, by the way.
1) To resuspend the spores, don’t shake or vortex the tube. Instead, GENTLY tip the tube back
and forth until the spores are suspended. If spores are in a bottle or flask, then you can GENTLY swirl
to resuspend the spores.
2) We dilute the spores 1:13 (1 ml spores into 12 ml Sterile water for injection) for spraying
rabbits. I would suggest taking spores not from the very top of the tube and adding them to water. To
mix the new suspension (about 1.3 X 10^9 per ml) gently tip or swirl the container.
3) Before spraying, gently tip or swirl the spore suspension before gently pouring into the collison.
If you have any questions, please call me at . If you are still having technical problems
with the spores you should get next Tuesday, please let me know. will come up the following
week (what day is best for you?) If you think my presence would be valuable, let me know, and I’ll also
come. Otherwise, it will just be her. (I don’t mind coming at all – I would just like my presence there to
serve a useful purpose, and not be just a warm body.)
– Bruce

From:
Sent: Friday, June 15, 2001 1:01 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: Spores and counting
Dr. Ivins,
A question on how you enumerate. Our SOPs say a plate should contain from
25-250 spores per plate (we do 5 plates per dilution). Do you have criteria
for rejecting low or high numbers? Say I get a plate that has 12 colonies
and all remaining plates are within the 25-250 range. Do you reject that
plate and average the 4 remaining, use all 5 and average, reject all 5 and
re-enumerate with 5 more etc. I ask this, because it could potentially be a
GLP issue. I apologize if this is any SOP that you have sent but
I have not seen them yet.
–
From: Ivins, Bruce E Dr USAMRIID
To:
Subject: RE: Spores and counting
Date: Friday, June 15, 2001 2:08:24 PM
We don’t have a specific number. When we are counting AGI’s, after plating, we put the samples back
into the cold until the next day. We examine all of the plates. If one group is contaminated or out of
range, we will go back and redilute and replate from the AGI sample. We have had to do that only a
handful of times out of thousands of samples. We usually do AGIs at 3 per set, 10-4 and 10-5 dilutions
(3 plates for each dilution). If you are getting low counts, you might do 10-3 and 10-4 dilutions. If we
get at least 2 of 3 readable plates, we go ahead and count the set and average the counts.
– Bruce

—–Original Message—–
From: ]
Sent: Friday, June 15, 2001 1:56 PM
To: ‘Ivins, Bruce E Dr USAMRIID’
Subject: RE: Spores and counting
How many have to be low or high before you reject the whole set?
—–Original Message—–
From: Ivins, Bruce E Dr USAMRIID [mailto:Bruce.Ivins@DET.AMEDD.ARMY.MIL]
Sent: Friday, June 15, 2001 1:20 PM
To: ‘
Subject: RE: Spores and counting
We count 15 – 150 colonies per plate. Because of the large size of the
colonies, it’s next to impossible to accurately count over 150 colonies on a
single plate. I have told many times that 15 – 150 is a more
realistic value than 25 – 250 or 30 to 300. If one plate is over count,
under count, or contaminated, we take note of it and disregard it in the
count and averaging.
– Bruce

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: FW: Spores
Date: Monday, June 18, 2001 11:18:57 AM
Attachments:
Hi,
I’m sending this to you again. I just wanted to make sure you received it. When I hear from you
about it, I’ll send it forward here. I think if you send us material about every 2-4 weeks, that would be
good. That will give us the time to purify each batch. Hope you enjoyed Annapolis! I thought there
were some good talks there.
– Bruce

From: Ivins, Bruce E Dr USAMRIID
To: ” Ivins, Bruce E Dr USAMRIID
Cc:
Subject: RE: Spores and counting
Date: Monday, June 18, 2001 1:33:51 PM
,
The spores were sent to this morning by Federal Express. Please let me know when you
get them. Also, keep me posted on your tests with them, so, if we need to, we can plan a trip up there
for next week. Remember, avoid vortexing and vigorously shaking them. If it seems as if the spores I
sent you are clumped a bit at the top, then take the spores you need from about midway down into the
suspension after you resuspend them.
– Bruce

The spores were sent to this morning by Federal Express. Please let me know when you
get them. Also, keep me posted on your tests with them, so, if we need to, we can plan a trip up there
for next week. Remember, avoid vortexing and vigorously shaking them. If it seems as if the spores I
sent you are clumped a bit at the top, then take the spores you need from about midway down into the
suspension after you resuspend them.
– Bruce

From: Ivins, Bruce E Dr USAMRIID
To:
Subject: FW: Spores and counting
Date: Monday, June 18, 2001 1:39:40 PM
Hi, If Battelle still has trouble with the new spores I sent them this week, I think that there will be
a trip up there next week, with definitely, probably – he wants to see the facilities where
the rPA studies will be done – me maybe, and you if you’d like. I’ll let you know how things go.
– Bruce

GAO: The FBI is withholding from production this referenced email that the FBI agent confirms she reportedly saw.

It would be far more fruitful if GAO were to focus its efforts on obtaining the contemporaneous documents from September and October 2001 relating to Dr. Ivins lab than to do a “gap assessment” of the science. We all heard the NPR Science Friday and the experts confirm that there is no “there there” with respect to a case against Dr. Ivins in particular. For example, has GAO interviewed Patricia Fellows, Dr. Ivins’ assistant? Does she now remember what happened to the apple laptop computer installed in the suite in summer 2001 that she had located for the 2002 audit and put on the form submitted for the audit?

For example, Dr. Fellows’ and Dr. Bassett’s records and recollections could confirm where the bacteremia work was done — where it was confirmed that what had killed the rabbits was anthrax.

DXersaid

“Cross-examination permits a criminal defendant to probe the analyst’s competence, training, judgment and incentives or pressure for manipulation of the results. Such questioning can expose flaws in the forensic testing process that undercut the reliability of such evidence …”
FBI Agent Ed Montooth and prosecutor Rachel Lieber say that they regret Dr. Ivins’ death because it would prevent him from proving Dr. Ivins’ guilt in court.

To the contrary, both are perfectly free to explain why the rabbit documents don’t fully explain his time in the BL-3 in early October. The word “rabbit” has never passed their lips.

For example, consider the study of bacteremia contemplated by the rabbit protocol that the FBI failed to produce. That’s where you draw infected blood to confirm the rabbit died from anthrax and not some other cause.

GAO: Did FBI give you the rabbit protocol about the experiment with the 52 rabbits that arrived the week of September 24th, 2001? If not, why not? Why wasn’t it uploaded among the thousands of pages they offered as relevant to Dr. Ivins’ guilt? What could be more relevant than the documents explaining how he spent his time?

DXersaid

While the prosecutors and investigators have never even mentioned the experiment with the 52 rabbits, a leading vaccine publication published the research. The results of Dr. Bruce Ivins experiment on those nights in the lab involving the rabbit and the test of the effect of formaldehyde was reported in VACCINE.